Inbred corn line NPID4466

ABSTRACT

Basically, this invention provides for an inbred corn line designated NPID4466, methods for producing a corn plant by crossing plants of the inbred line NPID4466 with plants of another corn plant. The invention relates to the various parts of inbred NPID4466 including culturable cells. This invention also relates to methods for introducing transgenic transgenes into inbred corn line NPID4466 and plants produced by said methods.

FIELD OF THE INVENTION

This invention is in the field of corn breeding. The invention relatesto a corn plant and its seed designated NPID4466, derivatives andhybrids thereof. This invention also is in the field of hybrid cornproduction employing the present inbred.

BACKGROUND OF THE INVENTION

Corn plants (Zea mays L.) can be self-pollinating or cross pollinating.Self pollination for extended time periods produces homozygosity atalmost all gene loci, forming a uniform population of true breedingprogeny. These are inbreds. Crossing two homozygous inbreds producesheterozygous gene loci in hybrid plants and seeds.

Corn plant breeding is a process to develop improved corn germplasm inan inbred or hybrid. Hybrids are developed with inbreds which aredeveloped by selecting corn lines and self pollinating these lines forseveral generations to develop homozygous pure inbred lines. Two inbredlines are crossed and hybrid seed is produced. One inbred is emasculatedand the pollen from the other inbred pollinates the emasculated inbred.The emasculated inbred, often referred to as the female, produces thehybrid seed F1. Emasculation of the inbred can be done by detasselingthe seed parent, or the inbred could have a male sterility factor whichwould eliminate the need to detassel the inbred.

Whether the seed producing plant is emasculated due to detasseling orCMS or transgenes, the seed produced by crossing two inbreds in thismanner is hybrid seed. This hybrid seed is F1 hybrid seed. The grainproduced by a plant grown from a F1 hybrid seed is referred to as F2 orgrain. Although, all F1 seed and plants, produced by this hybrid seedproduction system using the same two inbreds should be substantially thesame, all F2 grain produced from the F1 plant will be segregating cornmaterial.

The hybrid seed production produces hybrid seed which is heterozygous.The heterozygous results in hybrid plants, which are robust and vigorousplants. Inbreds on the other hand are mostly homozygous. Thishomozygosity renders the inbred lines less vigorous. Inbred seed can bedifficult to produce since the inbreeding process in corn linesdecreases the vigor. However, when two inbred lines are crossed, thehybrid plant evidences greatly increased vigor and seed yield comparedto open pollinated, segregating corn plants. An important consequence ofthe homozygosity and the homogeneity of the inbred corn lines is thatall hybrid seed produced from any cross of two such elite lines will bethe same hybrid seed and make the same hybrid plant. Thus the use ofinbreds makes hybrid seed which can be reproduced readily.

The ultimate objective of the commercial corn seed companies is toproduce high yielding, agronomically sound plants that perform well incertain regions or areas of the Corn Belt.

SUMMARY OF THE INVENTION

The present invention relates to an inbred corn line NPID4466.Specifically, this invention relates to plants and seeds of this line.Additionally, this relates to a method of producing from this inbred,hybrid seed corn and hybrid plants with seeds from such, hybrid seed.More particularly, this invention relates to the unique combination oftraits that combine in corn line NPID4466.

Generally then, broadly the present invention includes an inbred cornseed designated NPID4466. This seed produces a corn plant.

The invention also includes the tissue culture of regenerable cells ofNPID4466 wherein the cells of the tissue culture regenerates plantscapable of expressing the genotype of NPID4466. The tissue culture isselected from the group consisting of leaf, pollen, embryo, root, roottip, guard cell, ovule, seed, anther, silk, flower, kernel, ear, cob,husk and stalk, cell and protoplast thereof. The corn plant regeneratedfrom NPID4466 or any part thereof is included in the present invention.The present invention includes regenerated corn plants that are capableof expressing NPID4466's genotype, phenotype or mutants or variantsthereof.

The invention extends to hybrid seed produced by planting, inpollinating proximity which includes using preserved corn pollen asexplained in U.S. Pat. No. 5,596,838 to Greaves, seeds of corn inbredlines NPID4466 and another inbred line if preserved pollen is not used;cultivating corn plants resulting from said planting; preventing pollenproduction by the plants of one of the inbred lines if two are employed;allowing cross pollination to occur between said inbred lines; andharvesting seeds produced on plants of the selected inbred. The hybridseed produced by hybrid combination of plants of inbred corn seeddesignated NPID4466 and plants of another inbred line are a part of thepresent invention. This inventions scope covers hybrid plants and theplant parts including the grain and pollen grown from this hybrid seed.

The invention further includes a method of hybrid F1 production. A firstgeneration (F1) hybrid corn plant produced by the process of plantingseeds of corn inbred line NPID4466; cultivating corn plants resultingfrom said planting; permitting pollen from another inbred line to crosspollinate inbred line NPID4466; harvesting seeds produced on plants ofthe inbred; and growing a harvested seed are part of the method of thisinvention.

The present invention also encompasses a method of introducing at leastone targeted trait into corn inbred line comprising the steps of: (a)crossing plant grown from the present invention seed which is therecurrent parent, representative seed of which has been deposited, withthe donor plant of another corn line that comprises at least one targettrait selected from the group consisting of male sterility, herbicideresistance, insect resistance, disease resistance, amylose starch, andwaxy starch to produce F1 plants; (b) selecting from the F1 plants thathave at least one of the targeted traits, forming a pool of progenyplants with the targeted trait; (c) crossing the pool of progeny plantswith the present invention which is the recurrent parent to producebackcrossed progeny plants with the targeted trait; (d) selecting forbackcrossed progeny plants that have at least one of the target traitsand physiological and morphological characteristics of corn inbred lineof the recurrent parent, listed in Table 1 forming a pool of selectedbackcrossed progeny plants; and (e) crossing the selected backcrossedprogeny plants to the recurrent parent and selecting from the resultingplants for the targeted trait and physiological and morphologicalcharacteristics of corn inbred line of the recurrent parent, listed inTable 1 and reselecting from the pool of resulting plants and repeatingthe crossing to the recurrent parent and selecting step in succession toform a plant that comprises the desired trait and all of thephysiological and morphological characteristics of corn inbred line ofthe recurrent parent in the present invention listed in Table 1 asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

This method and the following method of introducing traits can be donewith less back crossing events if the trait and/or the genotype of thepresent invention are selected for or identified through the use ofmarkers. SSR, microsatellites, SNP and the like decrease the amount ofbreeding time required to locate a line with the desired trait or traitsand the characteristics of the present invention. Backcrossing in two oreven three traits (for example the glyphosate, Europe corn borer, cornrootworm resistant genes) is routinely done with the use of markerassisted breeding techniques. This introduction of transgenes ormutations into a corn line is often called single gene conversion.Although, presently more than one gene particularly transgenes ormutations which are readily tracked with markers can be moved during thesame “single gene conversion” process, resulting in a line with theaddition of more targeted traits than just the one, but still having thecharacteristics of the present invention plus those characteristicsadded by the targeted traits.

The method of introducing a desired trait into corn inbred linecomprising: (a) crossing plant grown from the present invention seed,representative seed of which has been deposited the recurrent parent,with plant of another corn line that comprises at least one target traitselected from the group consisting of nucleic acid encoding an enzymeselected from the group consisting of phytase, stearyl-ACP desaturase,fructosyltransferase, levansucrase, amylase, invertase and starchbranching enzyme, the donor parent to produce F1 plants; (b) selectingfor the targeted trait from the F1 plants, forming a pool of progenyplants; (c) crossing the progeny plants with the recurrent parent toproduce backcrossed progeny plants; (d) selecting for backcrossedprogeny plants that have at least one of the target trait andphysiological and morphological characteristics of corn inbred line ofthe present invention as listed in Table 1 forming a pool of backcrossedprogeny plants; and repeating a step of crossing the new pool with therecurrent parent and selecting for the targeted trait and the recurrentparents characteristics until the selected plant is essentially therecurrent parent with the targeted trait or targeted traits. Thisselection and crossing may take at least 4 backcrosses if markerassisted breeding is not employed.

The inbred line and seed of the present invention are employed to carrythe agronomic package into the hybrid. Additionally, the inbred line isoften carrying transgenes that are introduced in to the hybrid seed.

Likewise included is a first generation (F1) hybrid corn plant producedby the process of planting seeds of corn inbred line NPID4466;cultivating corn plants resulting from said planting; permitting pollenfrom inbred line NPID4466 to cross pollinate another inbred line;harvesting seeds produced on plants of the inbred; and growing a plantfrom such a harvested seed.

A number of different techniques exist which are designed to avoiddetasseling in corn hybrid production. Some examples are switchable malesterility, lethal genes in the pollen or anther, inducible malesterility, male sterility genes with chemical restorers. There arenumerous patented means of improving upon the hybrid production system.Some examples include U.S. Pat. No. 6,025,546, which relates to the useof tapetum-specific promoters and the barnase gene to produce malesterility; U.S. Pat. No. 6,627,799 relates to modifying stamen cells toprovide male sterility. Therefore, one aspect of the current inventionconcerns the present invention comprising one or more gene(s) capable ofrestoring male fertility to male-sterile corn inbreds or hybrids and/orgenes or traits to produce male sterility in corn inbreds or hybrids.

The inbred corn line NPID4466 and at least one transgenic gene adaptedto give NPID4466 additional and/or altered phenotypic traits are withinthe scope of the invention. Such transgenes are usually associated withregulatory elements (promoters, enhancers, terminators and the like).Presently, transgenes provide the invention with traits such as insectresistance, herbicide resistance, disease resistance increased ordeceased starch or sugars or oils, increased or decreased life cycle orother altered trait.

The present invention includes inbred corn line NPID4466 and at leastone transgenic gene adapted to give NPID4466 modified starch traits.Furthermore this invention includes the inbred corn line NPID4466 and atleast one mutant gene adapted to give modified starch, acid or oiltraits, i.e. amylase, waxy, amylose extender or amylose. The presentinvention includes the inbred corn line NPID4466 and at least onetransgenic gene: bacillus thuringiensis, the bar or pat gene encodingPhosphinothricin acetyl Transferase, Gdha gene, GOX, VIP, EPSP synthasegene, low phytic acid producing gene, and zein. The inbred corn lineNPID4466 and at least one transgenic gene useful as a selectable markeror a screenable marker is covered by the present invention.

A tissue culture of the regenerable cells of hybrid plants produced withuse of NPID4466 genetic material is covered by this invention. A tissueculture of the regenerable cells of the corn plant produced by themethod described above is also included.

DEFINITIONS

In the description and examples, which follow, a number of terms areused. In order to provide a clear and consistent understanding of thespecifications and claims, including the scope to be given such terms,the following definitions are provided.

ALLELE—Any alternative forms of sequence. Diploid cells carry twoalleles of the genetic sequence. These two sequence alleles correspondto the same locus (i.e. position) on homologous chromosomes.

ELITE INBRED—Corn plant that is substantially homozygous and whichcontributes useful agronomic and/or phenotypic qualities when used toproduce hybrids that are commercially acceptable.

GENE SILENCING—The loss or inhibition of the expression of a gene.

GENOTYPE—genetic makeup.

LINKAGE—Refers to a tendency of a segment of DNA on the same chromosometo not separate during meiosis of homologous chromosomes. Thus duringmeiosis this segment of DNA remains unbroken more often than expected bychance.

LINKAGE DISEQUILIBRIUM—Alleles tendency to remain in linked groups whensegregating from parents to progeny, than expected from chance.

LOCUS—A defined segment of DNA. This segment is often associated with anallele position on a chromosome.

PHENOTYPE—The detectable characteristics of the corn plant. Thesecharacteristics often are detections of the genotype/environmentinteraction.

PLANT—this includes reference to an immature or mature whole plant,including a plant that has been detasseled or from which seed or grainhas been removed. Seed or embryo that will produce the plant is alsoconsidered to be the plant.

Early Season Trait Codes

Emergence Rating (EMRGR): Recorded when 50% of the plots in the trialare at V1 (1 leaf collar) growth stage.

-   -   1=All plants have emerged and are uniform in size    -   3=All plants have emerged but are not completely uniform    -   5=Most plants have emerged with some just beginning to break the        soil surface, noticeable lack of uniformity    -   7=Less than 50% of the plants have emerged, and lack of        uniformity is very noticeable    -   9=A few plants have emerged but most remain under the soil        surface.        Seedling Growth (SVGRR or Vigor): Recorded between V3 and V5        (3-5 leaf stage) giving greatest weight to seedling plant size        and secondary weight to uniform growth.    -   1=Large plant size and uniform growth    -   3=Acceptable plant size and uniform growth    -   5=Acceptable plant size and might be a little non-uniform    -   7=Weak looking plants and non-uniform growth    -   9=Small plants with poor uniformity        Purpling (PRPLR): Emergence and/or early growth rating. Purpling        is more pronounced on the under sides of leaf blades especially        on midribs.    -   1=No plants showing purple color    -   3=30% plants showing purple color    -   5=50% plants showing purple color    -   7=70% plants showing purple color    -   9=90+% plants showing purple color        Herbicide Injury (HRBDR) List the herbicide type, which is being        rated. Then rated each hybrid/variety injury as indicated below.    -   1=No apparent reduction in biomass or other injury symptoms    -   5=Moderate reduction in biomass with some signs of sensitivity    -   9=Severe reduction in biomass with some mortality        Mid-Season Traits Codes        Heat Units to 50% Silk (HUPSN): Recorded the day when 50% of all        plants within a plot show 2 cm or more silk protruding from the        ear. Converted days to accumulated heat units from planting.        Heat units to 50% Pollen Shed (HUPSN): Recorded the day when 50%        of all plants within a plot are shedding pollen. Converted days        to accumulated heat units from planting.        Plant Height (PLHTN): After pollination, recorded average plant        height of each plot. Measured from ground to base of leaf noted.        Plant Ear Height (ERHTN): After pollination record average ear        height of each plot. Measure from ground to base of ear node        (shank). —for Inbreds        Plant Ear Height in CM: After pollination record average ear        height of each plot. Measure from ground to base of ear node        (shank). —for Hybrids        Root Lodging Early % (ERTLP): Early root lodging occurs up to        about two weeks after flowering and usually involves        goosenecking. The number of root lodged plants are counted and        converted to percentage.        Shed Duration (Shed Duration): Sum of daily heat units for days        when plants in the plot are actively shedding pollen.        Foliar Disease (LFDSR): Foliar disease ratings taken one month        before harvest through harvest. The predominant disease should        be listed in the trial information and individual hybrid ratings        should be given.    -   1=No lesions to two lesions per leaf.    -   3=A few scattered lesions on the leaf. About five to ten percent        of the leaf surface is affected.    -   5=A moderate number of lesions are on the leaf. About 15 to 20        percent of the leaf surface is affected.    -   7=Abundant lesions are on the leaf. About 30 to 40 percent of        the leaf surface is affected.    -   9=Highly abundant lesions (>50 percent) on the leaf. Lesions are        highly coalesced. Plants may be prematurely killed.        Data collection (as described above) on the following diseases:

Common Rust (CR) Eye Spot (ES) Gray Leaf Spot (GLS) Northern Corn LeafBlight (NCLB) Stewart's Bacterial Wilt (SBW) Southern Corn Leaf Blight(SCLB) Southern Rust (SR) Corn Virus Complex (CVC)Corn response to diseases can also be rated as:

-   -   R=Resistant=1 to 2 rating    -   MR=Moderately Resistant=3 to 4 rating    -   MS=Moderately Susceptible=5 to 6 rating    -   S=Susceptible=7 to 9 rating        Preharvest Trait Codes        Heat units to Black Layer (HUBLN): The day when 50% of all        plants within a plot reach black layer stage is recorded.        Convert days to accumulated heat units from planting.        Harvest Population (HAVPN): The number of plants in yield rows,        excluding tillers, in each plot are counted.        Barren Plants (BRRNP): The number of plants in yield rows having        no ears and/or abnormal ears with less than 50 kernels are        counted.        Dropped Ears (DROPP): The numbers of ears lying on the ground in        yield rows are counted.        Stalk Lodging % (STKLP): Stalk lodging will be reported as        number of plants broken below the ear without pushing, excluding        green snapped plants. The number of broken plants in yield rows        are counted and converted to percent.        Root Lodging Late % (LRTLP): Late root lodging can usually start        to occur about two weeks after flowering and involves lodging at        the base of the plant. Plants leaning at a 30-degree angle or        more from the vertical are considered lodged. The number of root        lodged plants in yield rows are counted and converted to        percent.        Push Test for Stalk and Root Quality on Erect Plants % (PSTSP or        PCT Push or % Pushtest): The push test is applied to trials with        approximately five percent or less average stalk lodging. Plants        are pushed that are not root lodged or broken prior to the push        test. Standing next to the plant, the hand is placed at the top        ear and pushed to arm's length. Push one of the border rows        (four-row small plot) into an adjacent plot border row. The        number of plants leaning at a 30-degree angle or more from the        vertical, including plants with broken stalks prior to pushing        are counted. Plants that have strong rinds that snap rather than        bend over easily are not counted. The goal of the push test is        to identify stalk rot and stalk lodging potential, NOT ECB        injury.        PUSXN: Push ten plants and enter the number of plants that do        not remain upright.        Intactness (INTLR):    -   1=Healthy appearance, tops unbroken    -   5=25% of tops broken    -   9=majority of tops broken        Plant Appearance (PLTAR): This is a visual rating based on        general plant appearance taking into account all factors of        intactness, pest, and disease pressure.    -   1=Complete plant with healthy appearance    -   5=plants look OK    -   9=Plants not acceptable        Green Snap (GRSNP or PCTGS or % GreenSnap): Counted the number        of plants in yield rows that snapped below the ear due to        brittleness associated with high winds.        Stay-green (STGRP): This is an assessment of the ability of a        grain hybrid to retain green color as maturity approaches (taken        near the time of black-layer) and should not be a reflection of        hybrid maturity or leaf disease. Recorded % of green tissue.        This may be listed as a Stay Green Rating instead of a        Percentage.        Stay Green Rating (STGRR): This is an assessment of the ability        of a grain hybrid to retain green color as maturity approached        (taken near the time of black layer or if major differences are        noted later). This rating should not be a reflection of the        hybrid maturity or leaf disease.    -   1=solid Green Plant    -   9=no green tissue        Ear/Kernel Rots (KRDSR): If ear or kernel rots are present, husk        ten consecutive ears in each plot and count the number that have        evidence of ear or kernel rots, multiply by 10, and round up to        the nearest rating as described below. Identify and record the        disease primarily responsible for the rot.    -   1=No rots, 0% of the ears infected.    -   3=Up to 10% of the ears infected.    -   5=11 to 20% of the ears infected.    -   7=21 to 35% of the ears infected.    -   9=36% or more of the ears infected.        Grain Quality (GRQUR): Taken on husked ears after black layer        stage. The kernel cap integrity and relative amount of soft        starch endosperm along the sides of kernels is rated.    -   1=smooth kernel caps and or 10% or less soft starch    -   3=slight kernel wrinkles and or 30% soft starch    -   7=moderate kernel wrinkles and or 70% soft starch    -   9=severe kernel wrinkled and or 90% or more soft starch        Preharvest Hybrid Characteristics        Ear Shape (DESHR): Slender, Semi-Blocky, Blocky    -   1=Blocky    -   5=Semi-blocky    -   9=Slender        Ear Type (EARFR): Fixed, Semi-Fixed, Flex    -   1=Flex    -   5=Semi-flex    -   9=Fixed        Husk Cover (HSKCR): Short, Medium, Long    -   1=Long    -   5=Medium    -   9=Short        Kernel Depth (KRLNR): Shallow, Medium, Deep    -   1=Deep    -   5=Medium    -   9=Short (shallow)        Shank Length (SHLNR): Short, Medium, Long    -   1=Short    -   5=Medium    -   9=Long        Kernel Row Number (KRRWN): The average number of kernel rows on        3 ears.        Cob diameter (COBDR): Cob diameter is to be taken with template.    -   1: small    -   5: Medium    -   9: Large        Harvest Trait Codes        Number of Rows Harvested (NRHAN)        Plot Width (RWIDN)        Plot Length (RLENN)        Yield Lb/Plot (YGSMN): bushels per acre adjusted to 15.5%        moisture        Test Weight (TSTWN or TWT): test weight at harvest in pounds per        bushel        Moisture % (MST_P): percent moisture or grain at harvest        Adjusted Yield in Bu/A (YBUAN)        Color Codes        Kernel Type: (KRTPN)    -   1) Dent    -   2) Flint        Endosperm Type: (KRTEN)    -   1) normal    -   2) amylose    -   3) waxy    -   4) other        Sterile Type (MSCT):    -   1) no if yes cytoplasm type then:    -   2) c-type    -   3) s-type        Anthocyanin of Brace Roots (PBRCC): the presence of color on 60%        of the brace roots during pollen shed.    -   1) Absent    -   2) Faint    -   3) Moderate    -   4) Dark    -   5) Brace Roots not present    -   6) Green    -   7) Red    -   8) Purple        Anther Color (ANTCC): at 50 percent pollen shed observe the        color of newly extruded anthers, pollen not yet shed    -   1) Yellow    -   2) Red    -   3) Pink    -   4) Purple        Glume Color (GLMCC): color of glumes prior to pollen shed    -   1) Red    -   2) Green        Silk Color (SLKCC): Taken at a late flowering stage when all        plants have fully extruded silk. Silks at least 2″ long but        still fresh.    -   1) Yellow    -   2) Pink    -   3) Red        Kernel Color (KERCC): the main color of the kernel from at least        three ears per ear family.    -   1) Yellow    -   2) White        Cob Color (COBCC): the main color of the cob after shelling from        at least three ears per ear family.    -   1) Red    -   2) Pink    -   3) White

ABR. Description Input Value EMRGN Final number of plants per plot #REGNN Region Developed: # 1. Northwest 2. Northcentral 3. Northeast 4.Southeast 5. Southcentral 6. Southwest 7. Other CRTYN Cross type: 1. sc2. dc 3. 3w # 4. msc 5. m3w 6. inbred 7. rel. line 8. other KRTPN Kerneltype: 1. sweet 2. dent # 3. flint 4. flour 5. pop 6. ornamental 7.pipecorn 8. other EMERN Days to Emergence EMERN #Days ERTLP % Rootlodging: (before anthesis): #% GRSNP % Brittle snapping: (beforeanthesis): #% TBANN Tassel branch angle of 2nd primary lateral degreebranch (at anthesis): HUPSN Heat units to 50% pollen shed: #HU (fromemergence) SLKCN Silk color: #/Munsell value HU5SN Heat units to 50%silk: (from emergence) #HU DSAZN Days to 50% silk in adapted zone: #DaysHU9PN Heat units to 90% pollen shed: #HU (from emergence) HU19N Heatunits from 10% to 90% pollen shed: #HU DA19N Days from 10% to 90% pollenshed: #Days LSPUR Leaf sheath pubescence # of second leaf above the ear(at anthesis) 1-9 (1 = none): ANGBN Angle between stalk and 2nd leafdegree above the ear (at anthesis): CR2LN Color of 2nd leaf above#/Munsell the ear (at anthesis): value GLCRN Glume Color: #/Munsellvalue GLCBN Glume color bars perpendicular # to their veins (glumebands): 1. absent 2. present ANTCN Anther color: #/Munsell value PLQURPollen Shed: 1-9 (0 = male sterile) # HU1PN Heat units to 10% pollenshed: #HU (from emergence) LAERN Number of leaves above the top earnode: # LTBRN Number of lateral tassel # branches that originate fromthe central spike: EARPN Number of ears per stalk: # APBRR Anthocyaninpigment of brace roots: 1. # absent 2. faint 3. moderate 4. dark TILLNNumber of tillers: # HSKCN Husk color 25 days after 50% #/Munsell silk:(fresh) value MLWVR Leaf marginal waves: 1-9 (1 = none) # LFLCR Leaflongitudinal creases: 1-9 (1 = none) # ERLLN Length of ear leaf at thetop ear node: #cm ERLWN Width of ear leaf at the top ear node at the #cmwidest point: PLHTN Plant height to tassel tip: #cm ERHCN Plant heightto the top ear node: #cm LTEIN Length of the internode between the ear#cm node and the node above: LTASN Length of the tassel from top leaf#cm collar to tassel tip: HSKDN Husk color 65 days after 50% silk: (dry)#/Munsell value DSGMN Days from 50% silk to 25% grain moisture #Days inadapted zone: SHLNN Shank length: #cm ERLNN Ear length: #cm ERDINDiameter of the ear at the midpoint: #mm EWGTN Weight of a husked ear:#gm KRRWR Kernel rows: 1. indistinct 2. distinct # KRNAR Kernel rowalignment: 1. straight 2. # slightly curved 3. curved ETAPR Eartaper: 1. slight 2. average 3. extreme # KRRWN Number of kernel rows: #COBCN Cob color: #/Munsell value HSKTR Husk tightness 65 days after 50%silk: 1-9 # (1 = loose) COBDN Diameter of the cob at the midpoint: #mmYBUAN Yield: #kg/ha KRTEN Endosperm type: 1. sweet 2. 3 extra sweet 3.normal 4. high amylose 5. waxy 6. high protein 7. high lysine 8. supersweet 9. high oil 10. other KRCLN Hard endosperm color: #/Munsell valueALECN Aleurone color: #/Munsell value ALCPR Aleurone color pattern: 1.homozygous # 2. segregating KRLNN Kernel length: #mm KRWDN Kernel width:#mm KRDPN Kernel thickness: #mm K1KHN 100 kernel weight: #gm HSKCR Huskextension: 1. short (ear # exposed) 2. medium (8 cm) 3. long (8-10 cm)4. very long (>10 cm) KRPRN % round kernels on 13/64 slotted screen: #%HEPSR Position of ear 65 days after # 50% silk: 1. upright 2. horizontal3. pendent STGRP Staygreen 65 days after anthesis: # 1-9 (1 = worst)DPOPP % dropped ears 65 days after anthesis: % LRTRP % root lodging 65days after anthesis: % HU25N Heat units to 25% grain moisture: (from #HUemergence) HUSGN Heat units from 50% silk to 25% grain #HU moisture inadapted zone:

DETAILED DESCRIPTION OF THE INVENTION

The inbred provides uniformity and stability within the limits ofenvironmental influence for traits as described in the VarietyDescription Information (Table 1) that follows.

Male sterility and/or CMS systems for corn parallel the CMS type systemsthat have been routinely used in hybrid production in sunflower.

To produce these types of hybrids, the companies must develop inbreds,which carry needed traits into the hybrid combination. Hybrids are notoften uniformly adapted for the entire Corn Belt, but most often arespecifically adapted for regions of the Corn Belt. Northern regions ofthe Corn Belt require shorter season hybrids than do southern regions ofthe Corn Belt. Hybrids that grow well in Colorado and Nebraska soils maynot flourish in richer Illinois and Iowa soils. Thus, a variety of majoragronomic traits is important in hybrid combination for the various CornBelt regions, and has an impact on hybrid performance.

Inbred line development and hybrid testing have been emphasized in thepast half-century in commercial corn production as a means to increasehybrid performance. Inbred development can be by pedigree selection,haploid/dihaploid production, and recurrent selection. Pedigreeselection can be the selection in an F2 population produced from aplanned cross of two genotypes (often elite inbred lines), or selectionof progeny of synthetic varieties, open pollinated, composite, orbackcrossed populations. This type of selection is effective for highlyinheritable traits, but other traits, for example, yield requiresreplicated test crosses at a variety of stages for accurate selection.

Corn breeders select for a variety of traits in inbreds that impacthybrid performance along with selecting for acceptable parental traits.Such traits include but are not limited to: yield potential in hybridcombination, dry down, maturity, grain moisture at harvest, greensnap,resistance to root lodging, resistance to stalk lodging, grain quality,disease and insect resistance, ear, and plant height. Additionally,hybrid performance will differ in different soil types such as lowlevels of organic matter, clay, sand, black, high pH, low pH; or indifferent environments such as wet environments, drought environments,and no tillage conditions. These traits appear to be governed by acomplex genetic system that makes selection and breeding of an inbredline extremely difficult. Even if an inbred in hybrid combination hasexcellent yield (a desired characteristic), it may not be useful becauseit fails to have acceptable parental traits such as seed yield, seedsize, pollen production, good silks, plant height, etc.

To illustrate the difficulty of breeding and, developing inbred lines,the following example is given. Two inbreds compared for similarity of29 traits differed significantly for 18 traits between the two lines. If18 simply inherited single gene traits were polymorphic with genefrequencies of 0.5 in the parental lines, and assuming independentsegregation (as would essentially be the case if each trait resided on adifferent chromosome arm), then the specific combination of these traitsas embodied in an inbred would only be expected to become fixed at arate of one in 262,144 possible homozygous genetic combinations.Selection of the specific inbred combination is also influenced by thespecific selection environment on many of these 18 traits which makesthe probability of obtaining this one inbred even more remote. Inaddition, most traits in the corn genome are regrettably not singledominant genes but are multi-genetic with additive gene action notdominant gene action. Thus, the general procedure of producing a nonsegregating F1 generation and self pollinating to produce a F2generation that segregates for traits and selecting progeny with thevisual traits desired does not easily lead to an useful inbred. Greatcare and breeder expertise must be used in selection of breedingmaterial to continue to increase yield and the agronomics of inbreds andresultant commercial hybrids.

Certain regions of the Corn Belt have specific difficulties that otherregions may not have. Thus the hybrids developed from the inbreds haveto have traits that overcome or at least minimize these regional growingproblems. Examples of these problems include in the eastern corn beltGray Leaf Spot, in the north cool temperatures during seedlingemergence, in the Nebraska region CLN (Corn Lethal Necrosis) and in thewest soil that has excessively high pH levels. The industry oftentargets inbreds that address these issues specifically forming nicheproducts. However, the aim of most large seed producers is to provide anumber of traits to each inbred so that the corresponding hybrid can beuseful in broader regions of the Corn Belt. The new biotechnologytechniques such as Microsatellites, RFLPs, RAPDs and the like haveprovided breeders with additional tools to accomplish these goals.

The inbred has been produced through a dihaploid system or isself-pollinated for a sufficient number of generations to give inbreduniformity. During plant selection in each generation, the uniformity ofplant type was selected to ensure homozygosity and phenotypic stability.The line has been increased in isolated farmland environments with dataon uniformity and agronomic traits being observed to assure uniformityand stability. No variant traits have been observed or are expected inNPID4466.

The best method of producing the invention is by planting the seed ofNPID4466 which is substantially homozygous and self-pollinating or sibpollinating the resultant plant in an isolated environment, andharvesting the resultant seed.

TABLE 1 NPID4466 VARIETY DESCRIPTION INFORMATION #1 Type: Dent #2 RegionBest Adapted: Broadly adapted *MG **Maturity Hybrid Group RangeRM***(estimate) 4 98-102 100 #3 Plant Traits Brace- Anther Glume SilkRoot Cob Kernel Color Color Color* Color Color Color NPID- Red/ GreenYellow Faint White Yellow 4466 purple NP2171 Red/ Green Green Faint toWhite Yellow purple moderate NP2391 Green Green/ Green Faint Red Yellowyellow Inbred disease resistance levels Goss' GLS NCLB Wilt Eyespot SCLBNPID4466 4 1 2 1 7 NP2171 3 2 1 2 4 NP2391 3 2 2 2 5 *MG = Maturitygroup **Maturity is the number of days from planting to physiologicalmaturity (planting to black layer) ***RM = relative maturity *Silk Colortaken at a late flowering stage when all plants have fully extrudedsilk; silks at least 2″ long but still fresh.

The data provided above is often a color. The Munsell code is areference book of color, which is known and used in the industry and bypersons with ordinary skill in the art of plant breeding. The purity andhomozygosity of inbred NPID4466 is constantly being tracked usingisozyme genotypes.

Isozyme Genotypes for NPID4466

Isozyme data were generated for inbred corn line NPID4466 according toprocedures known and published in the art. The data in Table 2 gives theelectrophoresis data on NPID4466 and two compared inbreds. The threeinbreds differ for electrophoresis results.

TABLE 2 ELECTROPHORESIS RESULTS FOR NPID4466 Inbred ACP1_T ACP4_T ADH1TIDH1T NPID4466 2 2 4 4 NP2171 4 4 4 4 NP2391 4 4 4 4 Inbred IDH2T MDH1TMDH2T MDH3T NPID4466 6 6 3 16  NP2171 6 6 3 16  NP2391 6 6 3 16  InbredMDH4T MDH5T MDH6T PGD1T NPID4466 12  12  Mm   3.8 NP2171 12  12  Mm  3.8 NP2391 12  12  Mm 2 Inbred PGD2T PGM1T PGM2T PHI1_T NPID4466 5 9 44 NP2171 5 9 4 4 NP2391 5 9 4 4

Table 3 shows a comparison between NPID4466 and two comparable inbreds.

NPID4466 has significantly more yield than NP2391, and similar yield toNP2171. The present invention does have significantly differentrequirements for heat units to reach 50% silking or pollen shed thaneither of the two compared inbreds.

TABLE 3 PAIRED INBRED COMPARISON DATA  HeatUnits  HeatUnits Plant EarShed Inbred Yield to P50 to S50  Height   Height   Duration  NPID4466 114.4 1280.0 1295.4 61.3 20.3 213.8 NP2171 101.0 1303.7 1319.0 57.0 21.6143.7 Diff 13.4 23.7 23.7 3.8 1.0 81.0 # Expts 5.0 31.0 31.0 9.0 9.0 3.0Prob 0.2 0.016** 0.056* 0.044** 0.7 0.063* NPID4466 114.4 1284.9 1296.161.3 20.3 213.8 NP2931 96.6 1410.1 1440.0 63.8 22.0 189.0 Diff 17.8125.2 143.3 2.4 1.7 24.8 # Expts 5.0 36.0 35.0 9.0 9.0 4.0 Prob 0.011**0.000*** 0.000*** 0.2 0.3 0.3 *.05 < Prob <= .10 **.01 < Prob <= .05***.00 < Prob <= .01

Table 4 shows the GCA (General Combining Ability) estimates of NPID4466compared with the GCA estimates of the other inbreds. The estimates showthe general combining ability is weighted by the number ofexperiment/location combinations in which the specific hybridcombination occurs. The interpretation of the data for all traits isthat a positive comparison is a practical advantage. A negativecomparison is a practical disadvantage. The general combining ability ofan inbred is clearly evidenced by the results of the general combiningability estimates. This data compares the inbred parent in a number ofhybrid combinations to a group of “checks”. The check data is from ourcompany's and other companies' hybrids which are commercial products andpre-commercial hybrids, which were grown in the same sets and locations.

TABLE 4 % Late % Early % Stay Test % Stalk % Push Root Root DroppedFinal Green Hybrid Parent1 Parent2 N Yield Moisture Weight Lodging TestLodging Lodging Ears Stand %  1 NPID4466 14 −12.7 1.4 0.9 −16.2 −3.0 0.1 2 NPID4466 15 −14.5 1.8 0.7 −1.6 −1.4 0.3  3 NPID4466 16 7.8 −1.3 −0.9−8.8 0.4 5.5 0.0 1.6  4 NPID4466 14 −12.2 −0.1 −0.2 −11.8 1.3 0.1  5NPID4466 15 0.1 −0.6 −0.6 −15.3 9.7 0.1 0.0 −1.4  6 NPID4466 15 −12.4−2.3 −0.8 3.3 0.3 −1.3  7 NPID4466 15 17.6 −0.7 −0.4 −3.3 10.0 0.1 0.00.0  8 NPID4466 15 15.4 −1.2 −0.6 −3.0 9.6 0.1 0.0 0.0  9 NPID4466 5−7.8 −1.8 −0.5 −42.5 −2.5 −2.2 10 NPID4466 16 11.6 −0.1 −0.1 7.4 8.3−0.5 11 NPID4466 16 9.6 0.7 0.1 −0.1 11.4 0.0 12 NPID4466 16 16.7 −0.9−0.8 2.2 9.9 0.0 13 NPID4466 16 13.7 0.0 −0.2 0.2 9.9 0.0 14 NPID4466 133.8 0.7 0.1 1.3 0.0 15 NPID4466 6 −2.3 −1.9 −0.6 −2.7 −6.8 0.7 16NPID4466 16 8.5 −0.8 −0.1 3.5 17.4 0.0 17 NPID4466 14 −30.8 −1.5 −0.28.9 21.7 0.1 0.1 18 NPID4466 16 16.3 −0.5 −0.4 5.6 −11.4 0.0 19 NPID446614 9.9 −0.4 −0.5 −9.7 −2.2 0.1 20 NPID4466 14 −1.4 −0.5 −0.8 −13.0 −1.0−0.3 21 NPID4466 13 11.9 −1.6 −0.8 −1.9 21.7 3.8 0.9 0.0 22 NPID4466 141.8 0.6 0.2 13.3 0.0 23 NPID4466 9 8.9 −1.7 −0.2 −7.9 0.0 24 NPID4466 11−2.2 −3.6 −1.5 −0.4 0.0 −9.7 −0.6 25 NPID4466 16 −1.0 −0.3 0.2 −11.9 1.4−7.6 0.1 26 NPID4466 8 8.5 0.0 0.0 −25.7 0.2 27 NPID4466 15 −6.8 1.3 0.6−25.4 1.4 −0.8 −0.8 28 NPID4466 16 −9.8 −1.0 −0.2 −20.1 0.7 7.6 −0.2 29NPID4466 13 −11.9 −0.4 0.1 −19.8 0.7 7.6 −2.0 30 NPID4466 15 2.0 0.2 0.0−11.1 0.8 −7.6 −0.8 31 NPID4466 43 −2.0 0.9 0.5 −3.9 −25.8 0.2 2.5 0.0−0.9 32 NPID4466 16 −5.6 −0.2 −0.2 0.9 0.1 −0.8 33 NPID4466 15 −12.0 0.0−0.3 −5.3 0.8 3.6 0.0 0.7 34 NPID4466 16 17.8 0.2 0.5 −0.4 −5.2 3.3 0.01.7 35 NPID4466 15 −5.7 −0.1 0.2 −5.6 −0.7 −1.4 36 NPID4466 16 26.6 −0.40.0 −3.4 −4.0 −2.6 0.0 1.7 37 NPID4466 42 0.2 0.5 0.1 −5.8 −12.5 3.3 0.0−2.2 38 NPID4466 15 −5.2 −0.1 −0.3 −1.0 0.3 −1.0 39 NPID4466 72 16.5−0.2 0.1 −4.8 −9.5 0.5 3.5 0.1 0.1 40 NPID4466 36 2.1 −0.2 0.1 −0.2−17.9 −1.3 1.4 −1.3 41 NPID4466 14 1.0 0.4 −0.3 −4.3 0.3 0.4 42 NPID446638 5.8 0.3 −0.1 −6.8 −0.4 −2.3 1.4 −0.2 43 NPID4466 38 2.4 0.0 −0.1 −5.69.6 1.0 −1.1 −0.3 44 NPID4466 15 4.4 0.2 −0.4 −0.3 0.3 −0.1 45 NPID446614 −8.8 −2.2 −1.1 1.6 −0.4 −1.9 46 NPID4466 15 0.3 0.5 0.0 −1.4 0.3 0.247 NPID4466 11 5.6 −3.3 −1.8 3.5 0.3 −2.2 48 NPID4466 16 1.3 −0.6 0.1−13.3 −3.7 0.0 0.1 49 NPID4466 13 −1.4 −1.1 −0.3 −15.9 0.8 −7.6 −0.6 50NPID4466 14 −4.0 1.9 0.9 −0.8 −3.0 0.3 51 NPID4466 44 14.2 −0.5 −0.4 1.6−5.7 0.6 9.2 0.0 −0.5 52 NPID4466 31 5.6 −1.9 −1.1 0.3 12.2 1.6 −0.2−5.6 53 NPID4466 16 −0.8 0.3 0.2 −8.7 3.9 0.0 54 NPID4466 15 2.4 −1.2−0.6 −0.5 −1.9 −0.6 55 NPID4466 14 −6.2 −0.3 −0.4 −2.3 0.3 −1.7 56NPID4466 15 −0.3 −0.9 −0.5 −4.8 −1.9 0.2 57 NPID4466 15 −5.7 0.1 −0.1−3.9 0.6 −0.8 58 NPID4466 39 2.1 0.2 0.7 −2.5 −15.7 1.0 −0.5 −0.7 59NPID4466 15 −1.6 0.2 −0.2 −0.8 2.1 0.1 60 NPID4466 12 −3.6 −0.1 −0.3−0.3 −1.0 −0.1 61 NPID4466 14 8.0 1.0 0.4 −5.8 −5.4 0.2 62 NPID4466 1620.1 −1.8 −0.9 −0.3 0.5 1.8 0.0 −0.2 63 NPID4466 44 8.6 −0.3 0.1 −5.94.3 4.3 −4.7 0.0 −0.2 64 NPID4466 28 4.7 −0.2 −0.2 3.3 7.0 −9.3 0.9 −0.565 NPID4466 15 −6.4 −0.1 0.2 −1.3 6.3 0.1 0.0 0.0 66 NPID4466 47 8.9 1.40.7 −5.6 −38.8 −1.4 −1.7 0.0 0.6 −13.9 67 NPID4466 16 −9.1 −0.2 −0.2−0.2 0.8 0.2 68 NPID4466 15 −5.9 0.6 0.2 1.3 2.3 0.2 69 NPID4466 16−12.2 1.0 0.6 −0.5 3.1 0.1 70 NPID4466 15 −6.0 0.6 0.2 −6.5 1.7 0.2 71NPID4466 14 −4.0 0.9 −0.1 −5.0 0.3 0.3 72 NPID4466 15 4.4 0.9 0.7 −23.41.4 −7.6 −0.1 73 NPID4466 16 3.3 2.2 1.3 −18.1 −57.4 7.6 −0.3 74NPID4466 15 3.3 0.2 0.2 −21.4 −3.6 −15.2 0.2 75 NPID4466 16 −14.2 0.00.3 −20.5 0.7 −7.6 0.2 76 NPID4466 31 11.1 −0.7 −0.6 −2.1 0.4 4.7 0.00.5 77 NPID4466 23 −8.1 −1.6 0.6 4.4 −8.1 4.2 0.1 78 NPID4466 15 6.7−2.9 −0.8 −5.3 0.8 7.6 0.2 79 NPID4466 14 2.5 −1.7 −0.9 −1.8 0.3 −1.7 80NPJD4466 15 7.3 −1.0 −0.8 −3.1 0.3 0.1 81 NPID4466 12 −3.2 1.0 0.4 −2.76.5 0.1 82 NPID4466 14 3.7 −1.9 −0.8 −4.0 −2.8 −1.0 83 NP1D4466 15 0.2−1.0 −0.9 3.5 0.3 −1.1 84 NPID4466 10 −13.7 −2.5 −1.6 1.4 6.5 0.4 85NPID4466 12 −0.7 2.1 1.7 −11.6 34.3 −1.3 −1.1 86 NPID4466 15 −5.3 −1.4−0.8 2.9 −0.4 −0.1 87 NPID4466 15 6.9 −0.7 −0.3 1.4 0.3 −0.3 88 NPID446612 −3.8 0.6 0.2 1.4 6.5 −0.1 89 NPID4466 12 6.9 0.6 0.1 −1.1 0.5 0.0 90NPID4466 16 4.5 −3.9 −1.8 2.2 3.3 9.8 0.0 91 NPID4466 16 −1.4 0.5 0.24.2 1.6 10.1 −0.8 XR= 1650 2.5 −0.3 −0.1 −4.4 −9.0 1.0 1.3 0.1 −0.4−13.9 XH= 91 1.1 −0.4 −0.2 −4.6 −9.0 1.1 1.0 0.0 −0.3 −13.9 XT= 11 6.50.1 0.1 −2.9 −11.1 0.9 −0.4 0.2 −0.3 −13.9 % Green % Emergence VigorHeatunits Heatunits Ear Plant Hybrid Parent1 Parent2 Snap Barren RatingRating to S50 to P50 Height Height  1 NPID4466 −16.4 −16.4  2 NPID44667.8 7.8 −3.4 −17.1  3 NPID4466 11.5 11.5 2.5 8.8  4 NPID4466 25.1 25.1 5 NPID4466 53.9 53.7 16.0 27.5  6 NPID4466 29.4 −3.1 14.4  7 NPID446653.9 53.7 1.0 22.5  8 NPID4466 53.9 53.7 21.0 17.5  9 NPID4466 0.0 10NPID4466 −0.4 −4.3 −3.6 −7.1 11 NPID4466 −0.4 −12.8 4.7 6.3 12 NPID4466−0.4 −8.8 11.4 1.3 13 NPID4466 −0.4 −12.8 8.1 −0.4 14 NPID4466 −0.4 0.34.7 4.6 15 NPID4466 0.0 16 NPID4466 −0.4 −24.8 −0.3 −3.8 17 NPID446612.4 10.4 −25.5 −17.5 18 NPID4466 1.1 −21.8 9.7 7.9 19 NPID4466 −16.4−16.4 20 NPID4466 11.6 11.6 21 NPID4466 32.4 30.4 19.5 22.5 22 NPID44665.5 −33.8 4.7 −3.8 23 NPID4466 24 NPID4466 0.0 6.7 −19.2 18.0 30.2 25NPID4466 0.0 −17.1 0.8 −5.2 26 NPID4466 −79.1 −11.7 2.3 27 NPID4466 0.0−52.1 13.3 7.3 28 NPID4466 0.0 −40.6 3.3 12.3 29 NPID4466 0.0 −20.1 −1.79.8 30 NPID4466 0.0 −29.1 −4.2 4.8 31 NPID4466 1.2 −0.4 −14.3 −14.2−10.6 4.0 32 NPID4466 21.3 21.3 5.8 0.2 33 NPID4466 0.0 0.0 2.5 −5.0 34NPID4466 0.0 0.0 −12.5 2.5 35 NPID4466 −6.4 −4.4 −15.0 8.6 36 NPID446622.0 43.0 −2.5 10.0 37 NPID4466 1.5 −1.3 −6.2 2.9 −8.4 11.9 38 NPID4466−14.6 −8.1 34.4 39 NPID4466 0.9 −0.1 −1.0 −5.2 5.8 3.4 11.2 40 NPID4466−0.1 0.9 −0.1 −9.3 −9.2 7.0 8.0 41 NPID4466 −2.6 −3.1 19.4 42 NPID44660.6 0.2 0.9 0.2 6.0 9.7 9.3 43 NPID4466 1.3 0.7 0.4 −8.8 −1.3 1.8 4.9 44NPID4466 −26.1 1.9 14.4 45 NPID4466 −2.6 1.9 −5.6 46 NPID4466 −14.6 −8.124.4 47 NPID4466 29.4 6.9 24.4 48 NPID4466 0.0 −17.1 10.8 7.3 49NPID4466 0.0 −55.1 −9.2 −2.7 50 NPID4466 10.8 32.7 51 NPID4466 0.0 0.1−27.7 −31.5 −17.1 7.8 52 NPID4466 1.0 1.3 −1.2 7.3 9.5 −9.1 15.3 53NPID4466 −19.9 −17.9 0.7 2.1 54 NPID4466 −14.6 −13.1 19.4 55 NPID4466−14.1 1.9 14.4 56 NPID4466 −14.6 1.9 4.4 57 NPID4466 7.8 7.8 −5.0 2.5 58NPID4466 −0.5 0.2 −1.1 −2.1 −1.6 −1.1 −3.3 59 NPID4466 38.7 40.7 −13.6−9.3 60 NPID4466 −0.9 8.3 11.9 61 NPID4466 1.6 17.9 62 NPID4466 −12.51.5 −13.8 −5.0 63 NPID4466 0.1 0.1 −26.5 −29.2 −2.6 14.3 64 NPID4466 7.56.5 9.5 17.5 65 NPID4466 73.9 73.7 21.0 27.5 66 NPID4466 0.3 0.2 0.2−0.2 −58.3 −43.8 3.4 1.8 67 NPID4466 −0.1 −9.5 1.2 −4.5 −10.5 68NPID4466 −1.1 −9.5 −15.8 10.5 9.5 69 NPID4466 8.1 10.1 −11.8 −7.9 70NPID4466 −0.3 −9.5 −15.8 −2.0 −8.0 71 NPID4466 17.9 1.9 14.4 72 NPID44660.0 −20.1 −6.7 2.3 73 NPID4466 0.0 27.4 0.8 14.8 74 NPID4466 0.0 −79.1−1.7 7.3 75 NPID4466 0.0 −4.1 10.8 −0.2 76 NPID4466 36.4 30.9 −1.5 −1.277 NPID4466 −0.3 −0.1 0.0 −4.8 3.6 6.1 4.5 78 NPID4466 0.0 −6.1 −6.7 2.379 NPID4466 29.4 −3.1 19.4 80 NPJD4466 35.4 16.9 39.4 81 NPID4466 1.18.3 1.9 82 NPID4466 8.1 10.1 1.4 −4.3 83 NP1D4466 17.4 11.9 24.4 84NPID4466 −2.9 −6.7 −18.1 85 NPID4466 0.0 86 NPID4466 −2.6 −3.1 19.4 87NPID4466 −14.6 1.9 24.4 88 NPID4466 −0.9 −14.2 −5.6 89 NPID4466 3.1 5.819.4 90 NPID4466 0.0 31.5 −15.0 25.0 91 NPID4466 −0.8 28.8 −4.8 −9.8 2.9XR= 0.3 0.2 0.4 −0.4 −7.4 −2.6 0.2 6.4 XH= 0.2 0.2 0.3 −0.3 −2.6 7.5 0.48.1 XT= 0.4 0.2 0.4 −0.1 −10.3 −8.1 0.1 6.6 XR = GCA Estimate: Weightedby Expt XH = GCA Estimate: Weighted by Parent2 XT = Same as XH but usingonly those parent2 with two years of data

Table 5 shows the inbred NPID4466 in hybrid combination, in comparisonwith two other hybrids, which are similar. When in this hybridcombination, the NPID4466 hybrid carries significantly more yield andmoisture in comparison to the other commercial hybrid 2. The commercialhybrid 3 and the present invention are also significantly different,with NPID4466 hybrid carrying significantly more yield and moisture thanhybrid 3.

TABLE 5 PAIRED HYBRID COMPARISON DATA Year Hybrid Yield Moist TWT PCTERLPCTSL Overall NPID4466 Hybrid 200.8 22.2 54.1 3.5 4.0 Hybrid 2 191.920.3 54.7 7.2 5.7 # Expts 46.0 49.0 21.0 4.0 12.0 Diff 8.9 2.0 0.6 3.71.7 Prob 0.023** 0.000*** 0.013** 0.5 0.6 Year Hybrid PCTPUSH PCTLRLStand PCTGS Emerge Overall NPID4466 Hybrid 21.7 3.9 102.9 1.3 5.1 Hybrid2 26.7 0.8 105.0 0.0 3.7 # Expts 6.0 7.0 51.0 2.0 15.0 Diff 5.0 3.1 2.01.3 1.3 Prob 0.8 0.2 0.2 0.3 0.001*** Year Hybrid Vigor HUS50 HUP50Pltht Earht Overall NPID4466 Hybrid 5.0 1257.0 1255.0 274.3 97.7 Hybrid2 3.5 1240.0 1240.0 261.7 99.9 # Expts 2.0 7.0 7.0 7.0 7.0 Diff 1.5 18.015.3 12.6 2.1 Prob 0.2 0.079* 0.2 0.004*** 0.7 Year Hybrid Yield MoistTWT PCTERL PCTSL Overall NPID4466 Hybrid 191.0 22.8 52.6 3.4 5.0 Hybrid3 177.8 20.2 54.3 0.0 3.4 # Expts 30.0 33.0 6.0 2.0 7.0 Diff 13.1 2.51.7 3.4 1.5 Prob 0.001*** 0.000*** 0.024** 0.5 0.5 Year Hybrid PCTPUSHPCTLRL Stand PCTGS Emerge Overall NPID4466 Hybrid 21.7 6.9 120.0 1.3 5.1Hybrid 3 46.7 7.4 123.3 0.5 3.3 # Expts 6.0 2.0 35.0 2.0 15.0 Diff 25.00.6 3.4 0.8 1.8 Prob 0.037** 0.9 0.3 0.1 0.000*** Year Hybrid VigorHUS50 HUP50 Pltht Earht Overall NPID4466 Hybrid 5.0 1245.0 1242.0 271.797.3 Hybrid 3 3.0 1249.0 1241.0 250.3 97.7 # Expts 2.0 5.0 5.0 6.0 6.0Diff 2.0 4.1 0.9 21.3 0.3 Prob 0.3 0.7 0.9 0.008*** 1.0 *.05 < Prob <=**.01 < Prob <= .05 ***.00 < Prob <= .01

Table 6 shows the yield response of NPID4466 in hybrid combination incomparison with two hybrids and the plants in the environment around itat the same location. The plot data for the present hybrid trendingabove hybrid 2 and hybrid 3 in moderate to high yielding environments.In the strip trial data there the NPID4466 hybrid is showingconsistently similar results in comparison to the environment level andslightly lower results than Hybrid 2 and Hybrid 3.

TABLE 6 YIELD RESPONSE Hybrid Error # Plots 75 100 125 150 175 200Research Plots NPID4466 Hybrid 22.6 47 67 97 128 158 189 219 Hybrid 221.5 1676 73 98 124 149 175 200 Hybrid 3 23.3 2251 93 113 134 154 175195 Strip Plots NPID4466 10.7 171 77 101 125 150 174 198 Hybrid 2 19.52071 84 107 130 153 176 199 Hybrid 3 18.3 2818 81 104 128 152 175 199

In the standard CMS system there are three different corn lines requiredto make the hybrid. First, there is a cytoplasmic male-sterile lineusually carrying the CMS or some other form of male sterility. This linewill be the seed producing parent line. Second, there must be a fertileinbred line that is the same or isogenic with the seed producing inbredparent but lacking the trait of male sterility. This is a maintainerline needed to make new inbred seed of the seed producing male sterileparent. Third there is a different inbred which is fertile, has normalcytoplasm and carries a fertility restoring gene. This line is calledthe restorer line in the CMS system. The CMS cytoplasm is inherited fromthe maternal parent (or the seed producing plant); therefore for thehybrid seed produced on such plant to be fertile the pollen used tofertilize this plant must carry the restorer gene. The positive aspectof this is that it allows hybrid seed to be produced without the needfor detasseling the seed parent. However, this system does requirebreeding of all three types of lines: 1) male sterile-to carry the CMS:2) the maintainer line; and, 3) the line carrying the fertility restorergene.

In some instances, sterile hybrids are produced and the pollen necessaryfor the formation of grain on these hybrids is supplied by interplantingof fertile inbreds in the field with the sterile hybrids.

A number of different inventions exist which are designed to avoiddetasseling in corn hybrid production. Some examples are switchable malesterility, lethal genes in the pollen or anther, inducible malesterility, male sterility genes with chemical restorers, sterility geneslinked with a parent. U.S. Pat. No. 6,025,546, relates to the use oftapetum-specific promoters and the barnase gene. U.S. Pat. No. 6,627,799relates to modifying stamen cells to provide male sterility. Therefore,one aspect of the current invention concerns the present inventioncomprising one or more gene(s) capable of restoring male fertility tomale-sterile corn inbreds or hybrids.

This invention also is directed to methods for producing a corn plant bycrossing a first parent corn plant with a second parent corn plantwherein the first or second parent corn plant is an inbred corn plantfrom the line NPID4466. Further, both first and second parent cornplants can come from the inbred corn line NPID4466 which produces a selfof the inbred invention. The present invention can be employed in avariety of breeding methods which can be selected depending on the modeof reproduction, the trait, and the condition of the germplasm. Thus,any breeding methods using the inbred corn line NPID4466 are part ofthis invention: selfing, backcrosses, hybrid production, and crosses topopulations, and haploid by such old and known methods of using KWSinducers lines, Krasnador inducers, stock six material that induceshaploids and anther culturing and the like.

The present invention may be useful as a male-sterile plant. Sterilitycan be produced by pulling or cutting tassels from the plant,detasseling, use of gametocides, or use of genetic material to renderthe plant sterile using a CMS type of genetic control or a nucleargenetic sterility. Male sterility is employed in a hybrid production byeliminating the pollen shed from the seed producing parent. The seedproducing parent is grown in isolation from other pollen sources exceptfor the pollen source which is the male fertile inbred, which serves asthe male parent in the hybrid. To facilitate pollination of the seedproducing (female) parent, the male fertile inbreds are planted mostoften in rows near the male sterile (female) inbred.

Methods for genetic male sterility are disclosed in EPO 89/3010153.8, WO90/08828, U.S. Pat. Nos. 4,654,465, 4,727,219, 3,861,709, 5,432,068 and3,710,511. Gametocides, some of which are taught in U.S. Pat. No.4,735,649 (incorporated by reference) can be employed to make the plantmale sterile. Gametocides including but not limited to glyphosate andits derivatives are chemicals or substances that negatively affect thepollen or at least the fertility of the pollen and provide malesterility to the seed producing parent.

Hybrid production employing any form of male sterility includingmechanical emasculation tends to have a small occurrence of selfpollinated female inbred seeds along with the intended F1 hybrid seeds.Great measures are taken to avoid the inbred seed production in a hybridseed production field; but inbred seed unfortunately does occur in F1seed production.

To detect inbred seed in a sample of putative hybrid seed the seeds canbe tested with molecular markers, but another method is to plant theseed. The seed planting process is an inbred capture process whichisolates inbred seed from hybrid F1 seed sources. This process forproducing selected inbred seed comprises planting a group of seedcomprising seed from a hybrid production, some of this seed may be seedof the hybrid parents. The inbred plants tend to be readily identifiablefrom the hybrid plants, the inbreds have a stunted appearance, i.e.shorter plant, smaller ear. Self pollination of the stunted plants grownfrom these identified inbred plants produces the female inbred seed orthe hybrid. The resultant plants are observed for size or they can betested by markers to identify any inbred plants. The identified inbredplants are selected and self-pollinated to form the inbred seed.

A number of well known methods can be employed to identify the genotypeof corn. The ability to understand the genotype of the present inventionincreases as the technology moves toward better markers for identifyingdifferent components within the corn genetic material. One of the oldestmethods is the use of isozymes which provides a generalized footprint ofthe material. Other markers adapted to provide a higher definitionprofile include Restriction Fragment Length Polymorphisms (RFLPs),Amplified Fragment Length Polymorphisms (AFLPs), Random AmplifiedPolymorphic DNAs (RAPDs), Polymerase Chain Reaction (there are differenttypes of primers or probes) (PCR), Microsattelites (SSRs), and SingleNucleotide Polymorphisms (SNPs), sequence selection markers just to lista few. The use of these marker techniques for gathering genotypeinformation from seeds and plants is well understood in the industry.The methods for using these techniques can be found in college textbookssuch as Breeding Field Crops, Milton et. al. Iowa State UniversityPress.

The marker profile of the inbred of this invention should be close tohomozygous for alleles. A marker profile produced with any of the locusidentifying systems known in the industry will identify a particularallele at a particular loci. A F1 hybrid made from the inbred of thisinvention will comprise a marker profile of the sum of both the profilesof its inbred parents. At each locus the allele for the presentinvention and the allele for the other inbred parent should be present.Thus the profile of the present invention will permit identification ofhybrids as containing the inbred parent of the present invention. Toidentify the female portion of any hybrid the hybrid seed material fromthe pericarp which is maternally inherited is employed in a markertechnique. The resultant profile is of the maternal parent. Thecomparison of this maternal profile with the hybrid profile will allowidentification of the paternal profile. The present invention includes acorn cell that is part of an inbred or hybrid plant which includes itsseed or plant part that has the marker profile of alleles of the presentinvention.

Marker systems are not just useful for identification of the presentinvention; they are also useful for breeding and trait conversiontechniques. Polymorphisms in corn permit the use of markers for linkageanalysis. If SSR are employed with flanking primers, the marker profilecan be developed with PCR and Southern Blots can often be eliminated.Use of flanking markers, PCR and amplification to genotype corn maturedof the material is well known by the industry. Primers for SSR markersand corn genome mapping information are publicly available through thehelp of the USDA at Corn GDB on the web.

Marker profiles of this invention can be employed to identifyessentially derived varieties or progeny developed with the inbred inits ancestry. This inbred may have progeny identified by having amolecular marker profile with genetic contribution of the present inbredinvention, as measured by either percent identity or percent similarity.

The present invention may have a new locus or trait introgressed throughdirect transformation, breeding methods including but not limited tobackcrossing or marker assisted breeding. A backcross conversion orlocus conversion both refer to a product of a backcrossing program.

The use of the present inbred as a recurrent parent in a breedingprogram it is often referred to as backcrossing. Backcrossing is oftenemployed to introgression a desired trait or trait(s), either transgenicor nontransgenic, into the recurrent parent. A plant with the trait orthe desired locus is crossed into recurrent corn parent usually in oneor more backcrosses. If markers are employed to assist in selection ofprogeny that have the desired trait and recurrent parent backgroundgenetics, then the number of backcrosses needed to recover the recurrentparent with the desired trait or locus can be relatively few two orthree. However, 3, 4, 5 or more backcrosses are often required toproduce the desired inbred with the gene or loci conversion in place.The number of backcrosses needed for a trait introgression is oftenlinked to the genetics of the line carrying the trait and the recurrentparent and the genetics of the trait. Multigenic traits, recessivealleles, unlinked traits play a role in the number of backcrosses thatmay be necessary to achieve the desired backcross conversion of theinbred.

In a book written by Hallauer entitled Corn and Corn Improvement,Sprague and Dudley, 3rd Ed. 1998 the basics of corn crossing techniquesalong with a number of other corn breeding methods such as recurrent,bulk or mass selection, pedigree breeding, open pollination breeding,marker assisted selection, double haploids development and selectionbreeding are taught. The ordinary corn breeder understands thesebreeding systems and how to apply such techniques to the presentinvention. Therefore, repetition of how to perform these breedingmethods is not listed within this application.

Dominant, single gene traits or traits with obvious phenotypic changesare particularly well managed in backcrossing programs. Prior totransformation and prior to markers, backcrossing was employed since the1950's to breed in identified corn traits.

The backcrossing program is more complicated when the trait is arecessive gene. To determine the presence of the recessive gene requiresthe use of some testing to determine if the trait I has beentransferred. Use of markers to detect the gene reduces the complexity oftrait identification in the progeny. A marker that is a SNP, specificfor the trait, can increase the efficiency and speed of tracking arecessive trait within a backcrossing program. Backcrossing of recessivetraits has allowed known mutation traits to be moved into more elitegermplasm. Mutations can be induced in germplasm by the plant breeder.Mutations can also result from plant or seed or pollen exposure totemperature alterations, culturing, radiation in various forms, chemicalmutagens like EMS and others. Some of the mutant genes which have beenidentified and introgressed into elite corn include the genotypes: waxy(wx), amylose extender (ae), dull (du), horny (h), shrunken (sh),brittle (bt), floury (fl), opaque (O), and sugary (su). Nomenclature formutant genes is based on the effect these mutant genes have on thephysical appearance and phenotype of the kernel. There are mutant geneswhich produce starch with markedly different functional properties eventhough the phenotypes of the seed and plant remain the same. Mutantsubspecies have generally been given a number after the named genotype,for example, sugary-1 (su1), sugary-2 (su2); shrunken 1 (sh1) andshrunken 2 (sh2). Traits such as Ht, waxy, brown mid-rib, amylose,amylose brown mid-rib, amylose extender (ae), opaque, dull, imazethapyrtolerant (IT or IR—designations for two different imazethapyr resistantgenes), sterility, fertility, phytic acid, NLB, SLB, and the like haveall been introgressed into elite inbreds through breeding programs. Thelast backcross generation is usually selfed if necessary to recover theinbred of interest with the introgressed trait.

All plants and plant cells produced using inbred corn line NPID4466 arewithin the scope of this invention. The invention encompasses the inbredcorn line used in crosses with other, different, corn inbreds to produce(F1) corn hybrid seeds and hybrid plants and the grain produced on thehybrid plant. This invention includes plant and plant cells, which upongrowth and differentiation produce corn plants having the physiologicaland morphological characteristics of the inbred line NPID4466.

This invention also includes transforming of introgressed transgenicgenes, or specific locus into the present invention. The prior art hasan extended list of transgenes, and of specific locus that carrydesirable traits. The transgenes that can be introgressed include butare not limited to insect resistant genes such as Corn Rootworm gene(s)in the event DAS-59122-7, Mir603 Modified Cry3A event, MON 89034, MON88017 Bacillus thuringiensis (Cry genes) Cry34/35Ab1, Cry1A. 105, POCry1F, Cry2Ab2, Cry1A, Cry1AB, Cry1Ac Cry3Bb1, or herbicide resistantgenes such as Pat gene or Bar gene, EPSP, the altered protoporphyrinogenoxidase (protox enzyme) U.S. Pat. Nos. 5,767,373, 6,282,837, WO01/12825, or disease resistant genes such as the Mosaic virus resistantgene, etc., or trait altering genes such as lignin genes, floweringgenes, oil modifying genes, senescence genes and the like.

The present invention also encompasses the addition of traits that focuson products or by products of the corn plant such as the sugars, oils,protein, ethanol, biomass and the like. The present invention caninclude a trait that forms an altered carbohydrate or altered starch. Analtered carbohydrate or altered starch can be formed by an introgressedgene(s) that affect the synthase, branching enzymes, pullanases,debranching enzymes, isoamylases, alpha amylases, beta amylases, AGP,ADP and other enzymes which effect the amylose, and or amylopectin ratioor content or the branching pattern of starch. The fatty acid modifyinggenes if introgressed into the present invention can also affect starchcontent. Additionally, introgressed genes that are associated with oreffect the starch and carbohydrates can be adapted so that the gene orits enzyme does not necessarily alter the form or formation of thestarch or carbohydrate of the seed or plant; instead the introgressedgene or its RNA, polypeptide, protein or enzyme adapted to degrade,alter, or otherwise change the formed starch or carbohydrate. Examplesof this technology are shown in U.S. Pat. Nos. 7,033,627, 5,714,474,5,543,570, 5,705,375, 7,102,057, which are incorporated by reference. Anexample of use of an alpha amylase adapted in this manner in corn isshown in U.S. Pat. No. 7,407,677 which is also incorporated byreference.

The methods and techniques for inserting, or producing and/oridentifying a mutation or making or reshuffling a transgene andintrogressing the trait or gene into the present invention throughbreeding, transformation, mutating and the like are well known andunderstood by those of ordinary skill in the art.

Various techniques for breeding, moving or altering genetic materialwithin or into the present invention (whether it is an inbred or inhybrid combination) are also known to those skilled in the art. Thesetechniques to list only a few are anther culturing, haploid/doublehaploid production, (stock six, which is a breeding/selection methodusing color markers and is a method that has been in use for forty yearsand is well known to those with skill in the art), transformation,irradiation to produce mutations, chemical or biological mutation agentsand a host of other methods are within the scope of the invention. Allparts of the NPID4466 plant including its plant cells produced using theinbred corn line are within the scope of this invention. The termtransgenic plant refers to plants having genetic sequences, which areintroduced into the genome of a plant by a transformation method and theprogeny thereof. Transformation methods are means for integrating newgenetic coding sequences into the plant's genome by the incorporation ofthese sequences into a plant through man's assistance, but not bybreeding practices. The transgene once introduced into plant materialand integrated stably can be moved into other germplasm by standardbreeding practices.

The recombinant DNA molecules of the invention can be introduced intothe plant cell in a number of art-recognized ways. uitable methods oftransforming plant cells include microinjection (Crossway et al.,BioTechniques 4:320-334 (1986)), electroporation (Riggs et al, Proc.Natl. Acad. Sci. USA 83:5602-5606 (1986)), agrobacterium mediatedtransformation (Hinchee et al., Biotechnology 6:915-921 (1988)), directgene transfer (Paszkowski et al., EMBO J. 3:2717-2722 (1984)), ballisticparticle acceleration using devices available from Agracetus, Inc.,Madison, Wis. and Dupont, Inc., Wilmington, Del. (see, for example,Sanford et al., U.S. Pat. No. 4,945,050; and McCabe et al.,Biotechnology 6:923-926 (1988)), protoplast transformation/regenerationmethods (see U.S. Pat. No. 5,350,689 issued Sep. 27, 1994 to Ciba-GeigyCorp.), Whiskers technology (See U.S. Pat. Nos. 5,464,765 and 5,302,523)and pollen transformation (see U.S. Pat. No. 5,629,183). Also see,Weissinger et al., Annual Rev. Genet. 22:421-477 (1988); Sanford et al.,Particulate Science and Technology 5:27-37 (1987)(onion); Christou etal., Plant Physiol. 87:671-674 (1988)(soybean); McCabe et al.,Bio/Technology 6:923-926 (1988)(soybean); Datta et al., Bio/Technology8:736-740 (1990)(rice); Klein et al., Proc. Natl. Acad. Sci. USA,85:4305-4309 (1988)(corn); Klein et al., Bio/Technology 6:559-563(1988)(corn); Klein et al., Plant Physiol. 91:440-444 (1988)(corn); Fromet al., Bio/Technology 8:833-839 (1990); Gordon-Kamm et al., Plant Cell2:603-618 (1990) (corn); and U.S. Pat. Nos. 5,591,616 and 5,679,558(Rice).

Vectors can include such items as: leader sequences, transitpolypeptides, promoters, terminators, genes, introns, marker genes, etc.The structures of the gene orientations can be sense, antisense, partialantisense, or partial sense: multiple gene copies can be used. Thetransgenic gene can come from various non-plant genes (such as;bacteria, yeast, animals, and viruses) along with being from plants.

The DNA is then transformed into the plant. A transgene introgressedinto this invention typically comprises a nucleotide sequence whoseexpression is responsible or contributes to the trait under the controlof a promoter appropriate for the expression of the nucleotide sequenceat the desired time in the desired tissue or part of the plant.Constitutive or inducible promoters are used. The transgene may alsocomprise other regulatory elements such as for example translationenhancers or termination signals. In an embodiment, the nucleotidesequence is the coding sequence of a gene and is transcribed andtranslated into a protein. In another embodiment, the nucleotidesequence encodes an antisense RNA, a sense RNA that is not translated oronly partially translated, a t-RNA, a r-RNA or a sn-RNA.

Where more than one trait is introgressed into this invention, it isthat the specific genes are all located at the same genomic locus in thedonor, non-recurrent parent, preferably, in the case of transgenes, aspart of a single DNA construct integrated into the donor's genome.Alternatively, if the genes are located at different genomic loci in thedonor, non-recurrent parent, backcrossing allows to recover all of themorphological and physiological characteristics of the invention inaddition to the multiple genes in the resulting corn inbred line.

In an embodiment, a transgene whose expression results or contributes toa desired trait to be transferred to the present invention comprises avirus resistance trait such as, for example, a MDMV strain B coatprotein gene whose expression confers resistance to mixed infections ofcorn dwarf mosaic virus and corn chlorotic mottle virus in transgeniccorn plants (Murry et al. Biotechnology (1993) 11:1559 64). In anotherembodiment, a transgene comprises a gene encoding an insecticidalprotein, such as, for example, a crystal protein of Bacillusthuringiensis or a vegetative insecticidal protein from Bacillus cereus,such as VIP3 (see for example, Estruch et al. Nat Biotechnol (1997)15:137 41). Also see, U.S. Pat. Nos. 5,877,012, 6,291,156; 6,107,2796,291,156 and 6,429,360. In another embodiment, an insecticidal geneintroduced into present invention is a Cry1Ab gene or a portion thereof,for example, introgressed into present invention from a corn linecomprising a Bt-11 event as described in U.S. Pat. No. 6,114,608, whichis incorporated herein by reference, or from a corn line comprising a176 event as described in Koziel et al. (1993) Biotechnology 11: 194200. In yet another embodiment, a transgene introgressed into presentinvention comprises a herbicide tolerance gene, for example, that isresistant to dicamba or a tolerance gene which provides of an alteredacetohydroxyacid synthase (AHAS) enzyme confers upon plants tolerance tovarious imidazolinone or sulfonamide herbicides (U.S. Pat. No.4,761,373). In another embodiment, a non-transgenic trait conferringtolerance to imidazolinones is introgressed into present invention (e.g.a “IT” or “IR” trait). U.S. Pat. No. 4,975,374, incorporated herein byreference, relates to plant cells and plants containing a gene encodinga mutant glutamine synthetase (GS) resistant to inhibition by herbicidesthat are known to inhibit GS, e.g. phosphinothricin and methioninesulfoximine. Also, expression of a Streptomyces bar gene encoding aphosphinothricin acetyl transferase in corn plants results in toleranceto the herbicide phosphinothricin or glufosinate (U.S. Pat. No.5,489,520). U.S. Pat. No. 5,013,659, which is incorporated herein byreference, is directed to plants that express a mutant acetolactatesynthase (ALS) that renders the plants resistant to inhibition bysulfonylurea herbicides. U.S. Pat. No. 5,162,602 discloses plantstolerant to inhibition by cyclohexanedione and aryloxyphenoxypropanoicacid herbicides. The tolerance is conferred by an altered acetylcoenzyme A carboxylase (ACCase). U.S. Pat. No. 5,554,798 disclosestransgenic glyphosate tolerant corn plants, which tolerance is conferredby an altered 5-enolpyruvyl-3-phosphoshikimate (EPSP) synthase gene.U.S. Pat. No. 5,804,425 discloses transgenic glyphosate tolerant cornplants, which tolerance is conferred by an EPSP synthase gene derivedfrom Agrobacterium tumefaciens CP-4 strain. Also, tolerance to aprotoporphyrinogen oxidase inhibitor is achieved by expression of atolerant protoporphyrinogen oxidase enzyme in plants (U.S. Pat. No.5,767,373). Another trait transferable to the present invention confersa safening effect or additional tolerance to an inhibitor of the enzymehydroxyphenylpyruvate dioxygenase (HPPD) and transgenes conferring suchtrait are, for example, described in WO 9638567, WO 9802562, WO 9923886,WO 9925842, WO 9749816, WO 9804685 and WO 9904021. All issued patentsreferred to herein are, in their entirety, expressly incorporated hereinby reference.

In an embodiment, a transgene transferred to present invention comprisesa gene conferring tolerance to a herbicide and at least anothernucleotide sequence encoding another trait, such as for example, aninsecticidal protein. Such combination of single gene traits is, forexample, a Cry1Ab gene and a bar gene. The introgression of a Bt11 eventinto a corn line, such as present invention, by backcrossing isexemplified in U.S. Pat. No. 6,114,608, and the present invention isdirected to methods of introgressing a Bt11 event into present inventionand to progeny thereof using for example the markers described in U.S.Pat. No. 6,114,608.

By way of example only, specific events (followed by their APHISpetition numbers) that can be introgressed into corn plants by backcrossbreeding techniques include the glyphosate tolerant event GA21 (97-09901p) or the glyphosate tolerant event NK603 (00-011-01p), the glyphosatetolerant/Lepidopteran insect resistant event MON 802 (96-31701p) Mon810,Lepidopteran insect resistant event DBT418 (96-29101p), male sterileevent MS3 (95-22801p), Lepidopteran insect resistant event Bt11(95-19501p), phosphinothricin tolerant event B16 (95-14501p),Lepidopteran insect resistant event MON 80100 (95-09301p) and MON 863(01-137-01p), phosphinothricin tolerant events T14, T25 (94-35701p),Lepidopteran insect resistant event 176 (94-31901p), Western cornrootworm (04-362-01p), the phosphinothricin tolerant and Lepidopteraninsect resistant event CBH-351 (92-265-01 p), and the transgenic cornevent designated 3272 taught in US application publication 20060230473(hereby incorporated by reference).

A further subject of the present invention is the plants which comprisetransformed cells, in particular the plants regenerated from transformedcells. Regeneration is effected by any suitable process, which dependson the nature of the species as described, for example, in thereferences hereinabove. Patents and patent applications which are citedin particular for the processes for transforming plant cells andregenerating plants are the following: U.S. Pat. Nos. 4,459,355,4,536,475, 5,464,763, 5,177,010, 5,187,073, EP 267,159, EP 604 662, EP672 752, U.S. Pat. Nos. 4,945,050, 5,036,006, 5,100,792, 5,371,014,5,478,744, 5,179,022, 5,565,346, 5,484,956, 5,508,468, 5,538,877,5,554,798, 5,489,520, 5,510,318, 5,204,253, 5,405,765, EP 442 174, EP486 233, EP 486 234, EP 539 563, EP 674 725, WO 91/02071 and WO95/06128.

The use of pollen, cotyledons, zygotic embryos, meristems and ovum asthe target tissue can eliminate the need for extensive tissue culturework. Generally, cells derived from meristematic tissue are useful. Themethod of transformation of meristematic cells of cereal is taught inthe PCT application WO96/04392. Any number of various cell lines,tissues, calli and plant parts can and have been transformed by thosehaving knowledge in the art. Methods of preparing callus or protoplastsfrom various plants are well known in the art and specific methods aredetailed in patents and references used by those skilled in the art.Cultures can be initiated from most of the above-identified tissue. Theonly true requirement of the transforming plant material is that it canultimately be used to form a transformed plant.

Duncan, Williams, Zehr, and Widholm, Planta (1985) 165:322 332 reflectsthat 97% of the plants cultured that produced callus were capable ofplant regeneration. Subsequent experiments with both inbreds and hybridsproduced 91% regenerable callus that produced plants. In a further studyin 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988), 7:262265 reports several media additions that enhance regenerability ofcallus of two inbred lines. Other published reports also indicated that“nontraditional” tissues are capable of producing somatic embryogenesisand plant regeneration. K. P. Rao, et al., Corn Genetics CooperationNewsletter, 60:64 65 (1986), refers to somatic embryogenesis from glumecallus cultures and B. V. Conger, et al., Plant Cell Reports, 6:345 347(1987) indicates somatic embryogenesis from the tissue cultures of cornleaf segments. Thus, it is clear from the literature that the state ofthe art is such that these methods of obtaining plants are, and were,“conventional” in the sense that they are routinely used and have a veryhigh rate of success.

Tissue culture procedures of corn are described in Green and Rhodes,“Plant Regeneration in Tissue Culture of Corn,” Corn for BiologicalResearch (Plant Molecular Biology Association, Charlottesville, Va.1982, at 367 372) and in Duncan, et al., “The Production of CallusCapable of Plant Regeneration from Immature Embryos of Numerous Zea maysGenotypes,” 165 Planta 322 332 (1985). Thus, another aspect of thisinvention is to provide cells that upon growth and differentiationproduce corn plants having the physiological and morphologicalcharacteristics of the present invention.

Corn is used as human food, livestock feed, and as raw material inindustry. Sweet corn kernels having a relative moisture of approximately72% are consumed by humans and may be processed by canning or freezing.The food uses of corn, in addition to human consumption of corn kernels,include both products of dry- and wet-milling industries. The principalproducts of corn dry milling are grits, meal and flour. The cornwet-milling industry can provide corn starch, corn syrups, and dextrosefor food use. Corn oil is recovered from corn germ, which is aby-product of both dry- and wet-milling industries.

Corn, including both grain and non-grain portions of the plant, is alsoused extensively as livestock feed, primarily for beef cattle, dairycattle, hogs, and poultry. Industrial uses of corn include production ofethanol, corn starch in the wet-milling industry and corn flour in thedry-milling industry. The industrial applications of corn starch andflour are based on functional properties, such as viscosity, filmformation, adhesive properties, and ability to suspend particles. Thecorn starch and flour have application in the paper and textileindustries. Other industrial uses include applications in adhesives,building materials, foundry binders, laundry starches, explosives,oil-well muds, and other mining applications. Plant parts other than thegrain of corn are also used in industry: for example, stalks and husksare made into paper and wallboard and cobs are used for fuel and to makecharcoal.

The seed of the present invention or of the present invention furthercomprising one or more single gene traits, the plant produced from theinbred seed, the hybrid corn plant produced from the crossing of theinbred, hybrid seed and various parts of the hybrid corn plant, can beutilized for human food, livestock feed, and as a raw material inindustry.

The present invention therefore also discloses an agricultural productcomprising a plant of the present invention or derived from a plant ofthe present invention. The present invention also discloses anindustrial product comprising a plant of the present invention orderived from a plant of the present invention. The present inventionfurther discloses methods of producing an agricultural or industrialproduct comprising planting seeds of the present invention, growingplant from such seeds, harvesting the plants and processing them toobtain an agricultural or industrial product.

A deposit of at least 2500 seeds of this invention will be maintained bySyngenta Seeds Inc. Access to this deposit will be available during thetendency of this application to the Commissioner of Patents andTrademarks and persons determined by the Commissioner to be entitledthereto upon request. All restrictions on availability to the public ofsuch material will be removed upon issuance of a granted patent of thisapplication by depositing at least 2500 seeds of this invention at theAmerican Type Culture Collection (ATCC), at 10801 University Boulevard,Manassas, Va. 20110. The ATCC number of the deposit is PTA-11893. Thedate of deposit was May 26, 2011 and the seed was tested on Jun. 12,2011 and found to be viable. The deposit of at least 2500 seeds will befrom inbred seed taken from the deposit maintained by Syngenta SeedsInc. The ATCC deposit will be maintained in that depository, which is apublic depository, for a period of 30 years, or 5 years after the lastrequest, or for the enforceable life of the patent, whichever is longer,and will be replaced if it becomes nonviable during that period.

Additional public information on patent variety protection may beavailable from the PVP Office, a division of the U.S. Government.

Accordingly, the present invention has been described with some degreeof particularity directed to the embodiment of the present invention. Itshould be appreciated, though that the present invention is defined bythe following claims construed in light of the prior art so thatmodifications or changes may be made to the embodiment of the presentinvention without departing from the inventive concepts containedherein.

1. A seed of maize variety NPID4466 a representative sample on depositin ATCC Accession Number PTA-11893.
 2. A maize plant of maize varietyNPID4466, a representative sample of seed of said maize plant on depositin ATCC Accession Number PTA-11893.
 3. An F1 hybrid maize seed producedby crossing the plant of maize variety NPID4466 according to claim 2with a different maize plant variety.
 4. A maize plant part of the plantof claim
 2. 5. A method of producing a maize seed comprising crossingthe maize plant of claim 2 with itself or a second maize plant.
 6. Amaize plant or its parts produced by growing the F1 hybrid maize seed ofclaim
 3. 7. A cell from a maize tissue culture of regenerable cells ofthe plant of claim
 2. 8. A method of producing a maize plant varietyNPID4466 with a desired trait comprising: (a) crossing the NPID4466plant according to claim 2 with a plant of another maize line thatcomprises the desired trait to produce F1 progeny plants, wherein thedesired trait is selected from the group consisting of waxy starch,amylase, male sterility, modified carbohydrates, modified corn fiber,modified fatty acids metabolism, modified fatty acids, herbicideresistance, insect resistance, bacterial disease resistance, fungaldisease resistance, and viral disease resistance; (b) selecting F1progeny plants that have the desired trait to produce selected F1progeny plants; (c) crossing the selected progeny plants with NPID4466plants to produce backcross progeny plants; (d) selecting for backcrossprogeny plants that have the desired trait to produce selected desiredprogeny plants; and (e) repeating steps (c) and (d) at least one moretime to produce plants that comprise the desired trait and all of thephysiological and morphological characteristics of maize plant varietyNPID4466 when grown in the same environmental conditions.
 9. A maizeseed from maize variety NPID4466 with the desired trait produced by theprocess of claim
 8. 10. A method of producing a plant product comprisingobtaining the plant of claim 6 or a part thereof and producing saidplant product therefrom.
 11. A process of producing maize seed with atrait of amylase conferred by a transgene, comprising crossing the firstparent maize plant according to claim 2 with a second parent maize plantwith said transgene, and harvesting the resultant seed.
 12. A maize seedproduced by the method of claim
 5. 13. The maize seed of claim 12,wherein the maize seed is hybrid seed.
 14. A method of producing a maizeplant variety NPID4466 comprising an added desired trait, the methodcomprising introducing a transgene or trait allele conferring thedesired trait into the plant of maize variety NPID4466 a representativesample of seed on deposit in ATCC Accession Number PTA-11893.
 15. Themethod of claim 14 wherein the desired trait is herbicide resistance,selected from the group consisting of: glyphosate, sulfonylurea,imidazolinione, dicamba, glufosinate, phenoxy proprionic acid,cycloshexone, traizine, benzonitrile and broxynil.
 16. A maize plantobtained by the method of claim 14, wherein said desired trait confers atrait selected from the group consisting of herbicide tolerance; insecttolerance; resistance to bacterial, fungal, nematode or viral disease;waxy starch; male sterility; restoration of male fertility; modifiedcarbohydrate metabolism or modified fatty acid metabolism.
 17. A methodof producing a maize plant derived from variety NPID4466, the methodcomprising the steps of (a) growing a progeny plant produced by crossingthe plant of claim 2 with a second maize plant; (b) crossing the progenyplant with itself or a different plant to produce a seed of a nextprogeny plant of a subsequent generation; (c) growing said next progenyplant and crossing the next progeny plant of the subsequent generationwith itself or a different plant; and (d) repeating crossing and growingsteps for an additional 0-5 generations to produce a maize plant derivedfrom the variety NPID4466.
 18. The method of claim 10 wherein the plantproduct is silage, starch, ethanol, oil, syrup, meal, amylase, andprotein.
 19. A method for developing maize derived plants from the plantof claim 2 comprising the steps of crossing the plant of claim 2 with atleast one other plant to form a plant population; breeding said plantpopulation with plant breeding techniques selected from the groupconsisting of recurrent selection, backcrossing, pedigree breeding,genetic marker enhanced selection, dihaploid, forming breeding progenyand selecting maize derived plants from said breeding progeny.